有斑百合多倍体诱导及鉴定

为了建立野生有斑百合多基因型种子离体多倍体诱导方法及快速、准确和高效的多倍体鉴定技术，本研究在离体培养条件下，以野生有斑百合预培养7 d的无菌萌动种子(2n=2x=24)为材料，用不同含量的秋水仙素溶液进行不同时间的诱变处理。以种子下胚轴明显膨大作为变异植株早期鉴定的标准，通过流式细胞仪检测细胞核DNA含量，并结合染色体计数法对诱导后的植株进行倍性鉴定，探讨在相同的种子萌动状态下，不同含量、不同浸泡时间的秋水仙素诱变效果，并初步建立了种子离体多倍体诱导和鉴定技术流程。结果表明:0.10%的秋水仙素溶液可有效地诱导野生有斑百合萌动种子的染色体加倍，其中以浸泡处理36 h的诱导效果最好，诱导率达44.43%；与二倍体植株相比，四倍体植株表现出叶片肥厚且增宽、叶色深绿、叶表面轻微褶皱、叶柄粗壮、生长缓慢等特征。研究还发现，植物外部形态变化不仅在二倍体和四倍体之间存在，而且在不同基因型的四倍体之间也存在差异。本研究利用下胚轴的膨大作为早期筛选多倍体的有效指标，同时结合直接测定细胞中DNA含量和计数染色体数可快速有效地培育出野生有斑百合同源四倍体植株。本研究建立的离体多倍体诱导流程除可应用于有斑百合种子多倍体的诱导外，也可为其他野生百合种子离体染色体加倍提供参考。 

Polyploidy induction and identification in Lilium concolor var. pulchellum

To establish a protocol of polyploid induction from multi-genotype seeds of wild L. concolor var. pulchellum in vitro, and develop a rapid, accurate and effective technique of ploidy confirmation, sterile in vitro germinating seeds obtained from 7 days' pre-cultivation was incubated in different concentrations of colchicine solution for different exposure times. Then their hypocotyl swollen was used as primary selection criteria of putative polyploids. The ploidy levels of putative polyploids were detected by flow cytometry combined with chromosome counts. The effects of different colchicine concentrations and exposure times on tetraploid production were compared. We have developed an seeds in vitro polyploid induction and identification protocol. The results indicated that the most efficient procedure for chromosome doubling was 0. 10% cholchine treating the seeds for 36 hours, induction rate was 44. 43%. The tetraploid plants were significantly different in color, thickness and surface of leaves and petiole as well as growth speed from diploid plants. Furthermore, the variations also happened within the different tetraploid genotype groups. Therefore, the technique of hypocotyl swollen of seeds combined with flow cytometry to detect polyploidy was efficient, this protocol provides a feasible method for inducing wild L. concolor var. pulchellum polyploids. Except for inducing L. concolor var. pulchellum polyploids, this in vitro polyploid induction process can also be used as a reference for doubling the chromosomes of other wild lily seeds.